Comprehensive screening of anabolic steroids, corticosteroids, and acidic drugs in horse urine by solid-phase extraction and liquid chromatography-mass spectrometry

J Chromatogr A. 2006 Jul 7;1120(1-2):38-53. doi: 10.1016/j.chroma.2006.03.089. Epub 2006 May 2.


This paper reports two highly efficient liquid chromatography-mass spectrometry (LC-MS) methods for the screening of anabolic steroids, corticosteroids, and acidic drugs for the purpose of doping control in equine sports. Sample extraction was performed using a mixed-mode C8-SCX solid-phase extraction (SPE) cartridge. The first eluted fraction (acidic/neutral fraction) was base-washed and the resulting organic extract was used for the screening of anabolic steroids and corticosteroids by LC-MS using multiple reaction monitoring (MRM) in the positive electrospray ionisation (ESI) mode. The remaining aqueous extract was re-adjusted to pH 6 and acidic drugs were recovered by liquid/liquid extraction. Detection was again achieved using LC-MRM but in the negative ESI mode. A total of 40 anabolic steroids and corticosteroids, and over 50 acidic drugs, including some cyclooxygenase-2 (COX-2) inhibitors, oxicams, anti-diabetics, sedatives, diuretics and Delta(9)-tetrahydro-11-norcannabinol-9-carboxylic acid, could be covered by the two LC-MS methods. Both methods utilized a high efficiency reversed-phase column (3.3 cm L x 2.1 mm I.D. with 3 microm particles) coupled with a fast-scanning triple-quadrupole mass spectrometer to achieve fast turnaround times. The overall turnaround times for both methods were 10 min, inclusive of post-run and equilibration times.

MeSH terms

  • Adrenal Cortex Hormones / administration & dosage
  • Adrenal Cortex Hormones / chemistry
  • Adrenal Cortex Hormones / urine*
  • Anabolic Agents / administration & dosage
  • Anabolic Agents / chemistry
  • Anabolic Agents / urine*
  • Animals
  • Chromatography, High Pressure Liquid / methods*
  • Doping in Sports
  • Horses
  • Molecular Structure
  • Reproducibility of Results
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • Steroids / administration & dosage
  • Steroids / chemistry
  • Steroids / urine
  • Substance Abuse Detection / methods*


  • Adrenal Cortex Hormones
  • Anabolic Agents
  • Steroids